Parotiditis virus fluorencent amplification detection reagent box and detection method
A mumps virus and detection kit technology, applied in fluorescence/phosphorescence, biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of reaction product contamination, false positives, high requirements, etc., and achieve simple and convenient sampling , convenient and accurate judgment, high sensitivity effect
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[0065] The steps of the TaqMan-MGB detection method are as follows:
[0066] (1) RNA extraction
[0067] Take 200 ul of the gargle solution and extract viral RNA (1 μg / μL) with the Reansy Mini Kit from QIAGEN, Germany.
[0068] (2) Fluorescent PCR amplification
[0069] Take the quantitative fluorescence amplification detection reagent to prepare the reaction solution, the reaction system is 25 μL each, including:
[0070] 5×RT-PCR buffer 5μL
[0071] Dimercaptothreitol 5mM
[0072] RNase inhibitor RNasin 0.8 unit / μL
[0073] Specific amplification upstream primer 0.88μM
[0074] Specific amplification downstream primer 0.88μM
[0075] Specific probe 0.28μM
[0076] The deoxynucleoside triphosphate mixture is:
[0077] dATP, dTTP, dCTP, dGTP each 200μM
[0078] Ampli TaqR DNA Polymerase 2.5 enzyme activity units / reaction
[0079] AMV reverse transcriptase 50 enzyme activity units / reaction
[0080] Analysis sample viral RNA (1μg / μL) 5μL
[0081] Make up to 25μL with...
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